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ELISA kit for porcine circovirus type 2 antibody detection

A porcine circovirus and type 2 antibody technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problem of poor binding between coated antigens and antibodies, many non-specific reactions, and is not suitable for rapid and large-scale detection in veterinary clinics And other issues

Inactive Publication Date: 2013-04-10
BEIJING DAWEIJIA BIOTECH SHARE CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the laboratory diagnostic methods of PCV2 mainly include histopathological detection, virus isolation and identification, indirect immunofluorescence technology, immunohistochemical technology, PCR technology, etc., but these technologies require high test conditions and professional skills, and the operation The process is cumbersome and not suitable for the needs of rapid detection and large-scale detection in veterinary clinics
The enzyme-linked immunosorbent assay (ELISA) method has the characteristics of simple operation, rapidity, and high sensitivity. Although there are already some porcine circovirus type 2 antibody detection ELISA kits at home and abroad, the indirect ELISA method is mostly used, and the coated Poor antigen-antibody binding performance, more non-specific reactions

Method used

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  • ELISA kit for porcine circovirus type 2 antibody detection
  • ELISA kit for porcine circovirus type 2 antibody detection
  • ELISA kit for porcine circovirus type 2 antibody detection

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Embodiment 1

[0017] The preparation and application method of embodiment 1 porcine circovirus type 2 antibody detection ELISA kit

[0018] 1. The preparation method of porcine circovirus type 2 antibody detection ELISA kit:

[0019] Antibody detection plate coating antigen preparation: using the PCV2-TJ (serial number AY181946) strain gene sequence in GenBank as a template, use the software Primer5.0 to design a pair of specific primers, upstream primer: 5'-GTGGTCTCCAGGTATGACGTATCCAAGGAGGC-3'; downstream Primer: 5'-GTCTCAAGATCTTCAAGGGTTAAGTGGGGGGT-3', FokI and SalI restriction sites were added to the 5' ends of the two primers, and the DNA product extracted from PCV-2-infected PK15 cell culture was used as a template to amplify by PCR. The full gene of the Cap protein of the laboratory-isolated strain PCV-2 was added. Through the designed FokI / SalI double restriction site, the target gene Cap was cloned into the red yeast secretion expression vector pP-secSUMO3, and transformed into Escher...

Embodiment 2

[0030] Example 2 The specificity, sensitivity and comparative test with similar products of porcine circovirus type 2 antibody detection ELISA kit

[0031] 1. specificity test: use test kit of the present invention to respectively to swine fever virus (CSFV) antibody-positive serum, porcine reproductive and respiratory syndrome virus (PRRSV) antibody-positive serum, porcine foot-and-mouth disease virus (FMDV) antibody-positive serum, pig pseudo Rabies virus (PRV) antibody positive serum, porcine parvovirus (PPV) antibody positive serum, porcine epidemic diarrhea virus (PEDV) antibody positive serum, porcine circovirus type I (PCV1) antibody positive serum, porcine circovirus type II (PCV2) antibody-positive serum is tested, and the result shows that the test result of PCV2 antibody-positive serum is positive, and all the other test results are negative. It shows that there is no cross-reaction between the antigen used in the kit of the present invention and the antibodies of o...

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Abstract

The invention discloses an ELISA (enzyme linked immunosorbent assay) kit for porcine circovirus type 2 (PVC2) antibody detection. The kit includes an antibody detection board, a sample diluting solution, an enzyme-labeled antigen, a washing solution, a substrate color developing solution, a stop solution, a positive serum and a negative serum. The Cap protein coated with antibody detection board of the kit is PCV2 eukaryotic expression nucleocapsid Cap protein, and the non-specific reactions are greatly reduced by using the Cap protein coupled enzyme-labeled antigen, so that the antibody detection accuracy is improved and the kit is more favorable for large-scale popularization and use.

Description

technical field [0001] The invention relates to an ELISA kit for detecting porcine circovirus type 2 antibody, which is mainly used in the detection of porcine circovirus type 2 antibody level, thereby judging the infection situation of porcine circovirus type 2 in pig farms and the immunity of vaccines Effect. Background technique [0002] Porcine circovirus type 2 (Porcine circovirus type 2, PCV2) is a non-enveloped single-stranded circular DNA virus with a particle diameter of about 17nm-20nm. It is one of the smallest animal viruses known so far. The mixed infection of PCV2 with other pathogens and the presence of other stress factors will induce the occurrence of porcine circovirus disease, which mainly includes multisystemic wasting syndrome in weaned piglets, porcine dermatitis nephrotic syndrome, porcine reproductive and respiratory syndrome, and neonatal piglets. Congenital tremor, porcine respiratory syndrome, porcine proliferative and necrotizing pneumonia, among...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
Inventor 董建辉
Owner BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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